CO2 and inorganic N supply modify competition for N between co-occurring grass plants, tree seedlings and soil microorganisms

Plant-plant and plant-soil interactions play a key role in determining plant community structure and ecosystem function. However, the effects of global change on the interplay between co-occurring plants and soil microbes in successional communities are poorly understood. In this study, we investigated competition for nitrogen (N) between soil microorganisms, grass plants and establishing tree seedlings under factorial carbon dioxide (CO₂) and N treatments. Fraxinus excelsior seedlings were germinated in the presence or absence of grass competition (Dactylis glomerata) at low (380 μmol mol⁻¹) or high (645 μmol mol⁻¹) CO₂ and at two levels of N nutrition in a mesocosm experiment. Pulse ¹⁵N labelling was used to examine N partitioning among plant and soil compartments. Dactylis exerted a strong negative effect on Fraxinus biomass, N capture and ¹⁵N recovery irrespective of N and CO₂ treatment. In contrast, the presence of Dactylis had a positive effect on the microbial N pool. Plant and soil responses to N treatment were of a greater magnitude compared with responses to elevated CO₂, but the pattern of Fraxinus- and microbial-N pool response to N and CO₂ varied depending on grass competition treatment. Within the Dactylis competition treatment, decreases in Fraxinus biomass in response to N were not mirrored by decreases in tree seedling N content, suggesting a shift from below- to above-ground competition. In the Dactylis-sown pots, ¹⁵N recovery could be ranked Dactylis > microbial pool > Fraxinus in all N and CO₂ treatment combinations. Inequalities between Fraxinus and soil microorganisms in terms of ¹⁵N recovery were exacerbated by N addition. Contrary to expectations, elevated CO₂ did not increase plant-microbe competition. Nevertheless, microbial ¹⁵N recovery showed a small positive increase in the high CO₂ treatment. Overall, elevated CO₂ and N supply did not interact on plant/soil N partitioning. Our data suggest that the competitive balance between establishing tree seedlings and grass plants in an undisturbed sward is relatively insensitive to CO₂ or N-induced modifications in N competition between plant and soil compartments.     

Transfer of N fixed by a legume tree to the associated grass in a tropical silvopastoral system

Below-ground transfer of nitrogen (N) fixed by legume trees to associated non-N₂-fixing crops has received little attention in agroforestry, although the importance of below-ground interactions is shown in other ecosystems. We used ¹⁵N natural abundance to estimate N transfer from the legume tree Gliricidia sepium (Jacq.) Kunth ex Walp. to C₄ grass Dichanthium aristatum (Poir.) C.E. Hubb. in a silvopastoral system, where N was recycled exclusively by below-ground processes and N₂ fixation by G. sepium was the sole N input to the system. Finding a suitable reference plant, a grass without contact with tree roots or litter, was problematic because tree roots invaded adjacent grass monocrop plots and soil isotopic signature in soil below distant grass monocrops differed significantly from the agroforestry plots. Thus, we used grass cultivated under greenhouse conditions in pots filled with agroforestry soil as the reference. A model of soil ¹⁵N fractionation during N mineralization was developed for testing the reliability of that estimate. Experimental and theoretical results indicated that 9 months after greenhouse transplanting, the percentage of fixed N in the grass decreased from 35% to <1%, due to N export in cut grass and dilution of fixed N with N taken up from the soil. The effect of soil ¹⁵N fractionation on the estimate of the reference value was negligible. This indicates that potted grass is a suitable reference N transfer studies using ¹⁵N natural abundance. About one third of N in field-grown grass was of atmospheric origin in agroforestry plots and in adjacent D. aristatum grassland invaded by G. sepium roots. The concentration of fixed N was correlated with fine root density of G. sepium but not with soil isotopic signature. This suggests a direct N transfer from trees to grass, e.g. via root exudates or common mycorrhizal networks.     

Estimating N rhizodeposition of grain legumes using a N in situ stem labelling method

Grain legumes in crop rotations cause significant increases in yield for succeeding non-legumes, which cannot be explained simply by the small effect that legumes have on the soil nitrogen balance, as found in the analysis of N in crop residues. Besides known positive non-N-effects, other effects, mainly rhizodeposition and its contribution to the N balance and nitrogen dynamics after harvesting the grain, are poorly understood. In this study, N rhizodeposition, defined as root-derived N in the soil after removal of visible roots, was measured in faba bean (Vicia faba L.), pea (Pisum sativum L.) and white lupin (Lupinus albus L.). In a pot experiment the legumes were pulse labelled in situ with ¹⁵N urea using a cotton wick method. About 84% of the applied ¹⁵N was recovered for the three legume species at maturity. The ¹⁵N was comparatively uniformly distributed among plant parts. The N rhizodeposition constituted 13% of total plant N for faba bean and pea and 16% for white lupin at maturity, about 80% of below ground plant N, respectively. Some 7% (lupin)-31% (pea) of the total N rhizodeposits were recovered as micro-roots by wet sieving (200 μm) the soil after all visible roots had been removed. Only 14-18% of the rhizodeposition N was found in the microbial biomass and a very small amount of 3-7% was found in the mineral N fraction. In pea, 48% and in lupin 72% of N rhizodeposits could not be recovered in the mentioned pools and a major part of the unrecovered N was probably immobilised in microbial residues. The results of this study clearly indicate that N rhizodeposition from grain legumes represent a significant pool for N balance and N dynamics in crop rotations.     

Nitrate removal from drained and reflooded fen soils affected by soil N transformation processes and plant uptake

Knowledge about nitrate transformation processes and how they are affected by different plants is essential in order to reduce the loss of valuable N fertiliser as well as to prevent environmental pollution due to nitrate leaching or N₂O emission after fertilisation or the reflooding of degraded fens with nitrate-containing municipal sewage. Therefore four microcosm ¹⁵N tracer experiments were performed to evaluate the effect of common wetland plants (Phalaris arundinacea, Phragmites australis) combined with different soil moisture conditions (from dry to reflooded) on nitrate turnover processes. At the end of experiment, the total formation of gaseous N compounds was calculated using the ¹⁵N balance method. In two experiments (wet and reflooded soil conditions) the N₂O and N₂ emissions were also directly determined.Our results show that in degraded fen soils, which process mainly takes place—denitrification or transformation into organic N compounds—is determined by the soil moisture conditions. Under dry soil moisture conditions (water filled pore space: 31%) up to 80% of the ¹⁵N nitrate added was transformed into organic N compounds. This transformation process is not affected by plant growth. Under reflooded conditions (water filled pore space: 100%), the total gaseous N losses were highest (77-95% of the ¹⁵N-nitrate added) and the transformation into organic N compounds was very low (1.8% of ¹⁵N nitrate added). Under almost all soil conditions plant growth reduced the N losses by 20-25% of the ¹⁵N nitrate added due to plant uptake. The N₂ emissions exceeded the N₂O emissions by a factor of 10-20 in planted soil, and as much as 30 in unplanted soil. In the treatments planted with Phragmites australis, N₂O emission was about two times higher than in the corresponding unplanted treatment. 15% of the N₂O and N₂ formed was transported via the Phragmites shoots from the soil into the atmosphere. By contrast, Phalaris arundinacea did not affect N₂O emissions and no emission via the shoots was observed.     

土壤C/N对苹果植株生长及氮素利用的影响

土壤C/N是土壤氮素循环的重要影响因素。本研究以2年生\"富士\"/平邑甜茶为试验材料, 应用¹⁵N示踪技术研究了不同土壤C/N[6.21(CK)、10、15、20、25、30、35和40]对苹果植株生长及氮素利用和损失的影响。结果表明: 随着土壤C/N比值的逐渐增大, 苹果新梢长度和植株鲜重均呈先升高后降低的变化趋势, C/N=15、20和25的3个处理苹果新梢长度和植株鲜重最大, 三者间无显著差异, 但均显著高于其他处理。不同C/N处理间植株15N利用率存在差异, 土壤C/N=25时, 植株¹⁵N利用率最大, 为22.87%, 与C/N=20的处理间无显著差异, 但两者均显著高于其他处理; 土壤C/N=40时, 植株¹⁵N利用率最低, 仅为15.43%, 低于CK处理的16.65%。土壤C/N处于15~25时, 植株吸收的氮素来自于肥料氮的比例较高; 而土壤C/N较低(<15)或太高(>25)时, 植株吸收的氮素来自于土壤氮的比例较高。土壤氮素残留量随土壤C/N的增大逐渐增加, C/N=40处理的土壤氮素残留量是CK的1.32倍。随着土壤C/N比值的逐渐增大, 肥料氮损失量呈先减少后增加的变化趋势, 以C/N=25时最少, 仅为施氮量的49.87%, 而对照最大, 为61.54%。因此, 综合土壤C/N对苹果植株生长及氮素平衡状况来看, 土壤C/N为15~25时, 能促进植株的生长发育, 降低氮肥损失, 提高肥料利用率。     

Effects of annual grass senescence on NH4 and N-glycine uptake by blue oak (Quercus douglasii) seedlings and soil microorganisms in California oak woodland

Annual grasses are stronger competitors for available soil N than blue oak seedlings and soil microorganisms. However, little is known about the dynamics of N competition during annual grass senescence. We conducted a field experiment in a California oak woodland to study effects of annual grass senescence on N uptake by grasses, blue oak seedlings and soil microorganisms. Labeled N was applied at the beginning of April, May and of June in the form of ¹⁵NH₄⁺ or ¹⁵N-glycine. Plants and soils were harvested after 5 days (¹⁵NH₄⁺ and ¹⁵N-glycine treatments) and after 26 days (¹⁵NH₄⁺ treatment only). We evaluated effects of N form, season and labeling period on N competition among oak seedlings, annual grasses and soil microorganisms. N forms did not affect competition among grasses, oak seedlings and soil microorganisms, but more ¹⁵N was incorporated into the soil organic N pool in the ¹⁵N-glycine treatments than in the ¹⁵NH₄⁺ treatments. There were no seasonal (May vs June) effects on ¹⁵N recovery in blue oak seedlings and soil microorganisms. Plant samples from April harvest were lost. In June, when grasses were senescing, more ¹⁵N was found in the soil inorganic pool than in May. Extremely dry soils in June may have limited inorganic N availability to oak seedlings and soil microorganisms. After 26-day labeling period, ¹⁵N recovery in blue oak seedlings and the soil organic N pool significantly increased, while ¹⁵N recovery in both the soil microbial and inorganic N pools decreased compared to the 5-day labeling period. Although blue oak seedling biomass changed little from early May to late June, N concentrations in oak roots increased 53%. In contrast, annual grass biomass peaked in May, and then decreased rapidly. Our results suggest that blue oak seedlings and annual grasses have different temporal competitive abilities. Blue oak seedlings appear to have a long-term strategy for N competition. Blue oaks take up N slowly but steadily, increasing N uptake from 5 to 26 days. This extended time period has a greater positive effect on N uptake than does reduced grass uptake caused by senescence.     

Soil organic N dynamics and stand quality in Pinus radiata pinewoods of the temperate humid region

Soil organic-N dynamics, its controlling factors and its relationships with stand quality were studied in the 0-15 cm soil layer of 24 pinewoods with contrasting age, productivity and parent material (granite; acid schists), searching for N variables useful to predict stand growth and site quality. No significant differences were found between young and old stands for any of the N variables considered, nor two- or three-order interactions among stand age, site quality and parent material. The soil total-N content, which was correlated positively with the Al oxides content (a soil organic matter (SOM) stabilizing agent), did not vary significantly according to parent material, but it was lower (P≤0.02) in stands with high than with low site index (2.68±1.11 and 3.97±1.13 g N kg⁻¹ soil, respectively). The soil δ¹⁵N ranged from +3.5 to +6.5 δ, without significant differences among stand groups, and it was negatively correlated with water holding capacity, exchangeable bases, Al oxides and N content, suggesting that: (i) N losses by NO₃⁻ leaching are the most important controlling factor of δ¹⁵N in these temperate humid region soils; and (ii) soil N richness is related with limited N losses, which discriminate against ¹⁵N. At any incubation time, no significant differences were found in soil inorganic-N content among stand groups (7.78±4.57, 39.33±16.20 and 67.80±26.50 mg N kg⁻¹ soil at 0, 42 and 84 d, respectively). During the incubation, the relative importance of ammonification decreased and that of the nitrification increased. The net N mineralization rate (NNMR, in percentage of organic N) was significantly higher in granite than in schists soils at both 42 d (1.24±0.34 and 0.75±0.37%, respectively) and 84 d (2.18±0.56 and 1.53±0.66%, respectively). In high quality pinewoods, the NNMR at 42 and 84 d (1.16±0.45 and 2.12±0.79%, respectively) were significantly higher than in low quality stands (0.83±0.35 and 1.59±0.45%, respectively). This result, together with those on soil total-N and inorganic-N supply, suggests that soil N dynamics in low and high quality stands are different: in the former there is a bigger N pool with a slower turnover, whereas in the latter there is a smaller N pool with a faster turnover, both factors being nearly compensated, making the soil available N supply in both types of stand similar. After 42 and 84 d of incubation, the NNMR and the nitrification rates were higher in the coarse textured soils, likely due to the low physical and chemical protection of their SOM; both rates were positively correlated with available P, exchangeable K⁺ and CEC base saturation, suggesting strong relationships among the availabilities of the main plant nutrients, and they increased with SOM quality (low C-to-N ratio). The strong negative correlation of site index with soil total-N (r=−0.707; P≤0.005), and its positive correlations with NNMR after 42 and 84 d of incubation, suggested that site quality and potential productivity are closely related to soil organic-N dynamics. Half of the site index variation in the stands studied could be predicted with a cheap and easy analysis of soil N content, the prediction being slightly improved if soil δ¹⁵N is included and, more significantly, by including N mineralization measurements.     

Elevated CO2, but not defoliation, enhances N cycling and increases short-term soil N immobilization regardless of N addition in a semiarid grassland

Elevated CO₂ and defoliation effects on nitrogen (N) cycling in rangeland soils remain poorly understood. Here we tested whether effects of elevated CO₂ (720 μl L⁻¹) and defoliation (clipping to 2.5 cm height) on N cycling depended on soil N availability (addition of 1 vs. 11 g N m⁻²) in intact mesocosms extracted from a semiarid grassland. Mesocosms were kept inside growth chambers for one growing season, and the experiment was repeated the next year. We added ¹⁵N (1 g m⁻²) to all mesocosms at the start of the growing season. We measured total N and ¹⁵N in plant, soil inorganic, microbial and soil organic pools at different times of the growing season. We combined the plant, soil inorganic, and microbial N pools into one pool (PIM-N pool) to separate biotic + inorganic from abiotic N residing in soil organic matter (SOM). With the ¹⁵N measurements we were then able to calculate transfer rates of N from the active PIM-N pool into SOM (soil N immobilization) and vice versa (soil N mobilization) throughout the growing season. We observed significant interactive effects of elevated CO₂ with N addition and defoliation with N addition on soil N mobilization and immobilization. However, no interactive effects were observed for net transfer rates. Net N transfer from the PIM-N pool into SOM increased under elevated CO₂, but was unaffected by defoliation. Elevated CO₂ and defoliation effects on the net transfer of N into SOM may not depend on soil N availability in semiarid grasslands, but may depend on the balance of root litter production affecting soil N immobilization and root exudation affecting soil N mobilization. We observed no interactive effects of elevated CO₂ with defoliation. We conclude that elevated CO₂, but not defoliation, may limit plant productivity in the long-term through increased soil N immobilization.     

Impacts of earthworms and arbuscular mycorrhizal fungi (Glomus intraradices) on plant performance are not interrelated

Earthworms and arbuscular mycorrhizal fungi (AMF) might interactively impact plant productivity; however, previous studies reported inconsistent results. We set up a three-factorial greenhouse experiment to study the effects of earthworms (Aporrectodea caliginosa Savigny and Lumbricus terrestris L.) and AMF (Glomus intraradices N.C. Schenck & G.S. Sm.) on the performance (productivity and shoot nutrient content) of plant species (Lolium perenne L., Trifolium pratense L. and Plantago lanceolata L.) belonging to the three functional groups grasses, legumes and herbs, respectively. Further, we investigated earthworm performance and plant root mycorrhization as affected by the treatments. Our results accentuate the importance of root derived resources for earthworm performance since earthworm weight (A. caliginosa and L. terrestris) and survival (L. terrestris) were significantly lower in microcosms containing P. lanceolata than in those containing T. pratense. However, earthworm performance was not affected by AMF, and plant root mycorrhization was not modified by earthworms. Although AMF effectively competed with T. pratense for soil N (as indicated by δ¹⁵N analysis), AMF enhanced the productivity of T. pratense considerably by improving P availability. Remarkably, we found no evidence for interactive effects of earthworms and AMF on the performance of the plant species studied. This suggests that interactions between earthworms and AMF likely are of minor importance.     

Temperature sensitivity of soil respiration is affected by prevailing climatic conditions and soil organic carbon content: A trans-China based case study

Understanding the spatial variation of temperature sensitivity (i.e. Q₁₀) of soil respiration (R_s) and its controlling factors, is critical to improve the precision of carbon budget estimations at regional scales. In this study, data from 2-3 continuous years of R_s measurements over 15 ecosystems of ChinaFLUX were summarized to analyze the response of R_s to soil temperature. Moreover, we improved our dataset by collecting previously published Q₁₀ values from 34 ecosystems in China. The ecosystems studied were located in the main climatic zones of China, spanning from alpine via temperate to tropical. Spatial variations of Q₁₀ and its controlling factors were analyzed. The results showed that soil temperature at a 5 cm depth satisfactorily explained the seasonal variations in R_s of the 15 ChinaFLUX ecosystems (R² varying from 0.37 to 0.83). Based on the overall data, the Q₁₀ values of R_s in China ranged from 1.28 to 4.75. The spatial variations in Q₁₀ were primarily determined by soil temperature during measurement periods, soil organic carbon (SOC) content, and ecosystem type. Ecosystems in colder regions and with higher SOC content had relatively higher Q₁₀ values. Moreover, ecosystems of different vegetation types showed different Q₁₀ values. A temperature- and SOC-dependent function for Q₁₀ is suggested, which could be a valuable reference for improving the regional-scale models of R_s and ecosystem carbon cycles.     

Impacts of different N management regimes on nitrifier and denitrifier communities and N cycling in soil microenvironments

Real-time quantitative PCR assays, targeting part of the ammonia monooxygenase (amoA), nitrous oxide reductase (nosZ), and 16S rRNA genes were coupled with ¹⁵N pool dilution techniques to investigate the effects of long-term agricultural management practices on potential gross N mineralization and nitrification rates, as well as ammonia-oxidizing bacteria (AOB), denitrifier, and total bacterial community sizes within different soil microenvironments. Three soil microenvironments [coarse particulate organic matter (cPOM; >250 μm), microaggregate (53-250 μm), and silt-and-clay fraction (<53 μm)] were physically isolated from soil samples collected across the cropping season from conventional, low-input, and organic maize-tomato systems (Zea mays L.-Lycopersicum esculentum L.). We hypothesized that (i) the higher N inputs and soil N content of the organic system foster larger AOB and denitrifier communities than in the conventional and low-input systems, (ii) differences in potential gross N mineralization and nitrification rates across the systems correspond with AOB and denitrifier abundances, and (iii) amoA, nosZ, and 16S rRNA gene abundances are higher in the microaggregates than in the cPOM and silt-and-clay microenvironments. Despite 13 years of different soil management and greater soil C and N content in the organic compared to the conventional and low-input systems, total bacterial communities within the whole soil were similar in size across the three systems (∼5.15 × 10⁸ copies g⁻¹ soil). However, amoA gene densities were ∼2 times higher in the organic (1.75 × 10⁸ copies g⁻¹ soil) than the other systems at the start of the season and nosZ gene abundances were ∼2 times greater in the conventional (7.65 × 10⁷ copies g⁻¹ soil) than in the other systems by the end of the season. Because organic management did not consistently lead to larger AOB and denitrifier communities than the other two systems, our first hypothesis was not corroborated. Our second hypothesis was also not corroborated because canonical correspondence analyses revealed that AOB and denitrifier abundances were decoupled from potential gross N mineralization and nitrification rates and from inorganic N concentrations. Our third hypothesis was supported by the overall larger nitrifier, denitrifier, and total bacterial communities measured in the soil microaggregates compared to the cPOM and silt-and-clay. These results suggest that the microaggregates are microenvironments that preferentially stabilize C, and concomitantly promote the growth of nitrifier and denitrifier communities, thereby serving as potential hotspots for N₂O losses.     

Changes in some enzyme activities and the nicotinamide nucleotide coenzymes in barley seedlings during vernalization

Respiratory metabolism was investigated by determining the following enzyme activities and nicotinamide nucleotide coenzyme contents in barley shoots (Hordeum vulgare L., var. Akashinriki) during vernalization : glucose-6-phosphate dehydrogenase, phosphoglucoisomerase, enolase, isocitrate dehydrogenase, malate dehydrogenase, and glutamic dehydrogenase. The activities of glucose-6-phosphate de hydrogenase and phosphoglucoisomerase in vernalized shoots were slightly lower than those in non-vernalized samples. Malate dehydrogenase activity increased gradually in vernalized shoots and was always higher than in non-vernalized shoots. Activation was found in enolase, isocitrate dehydrogenase and glutamic dehydrogenase after a long vernalization treatment.

The contents of total nicotinamide nucleotide coenzymes increased during the early germination stage in both treatments, then decreased gradually in the succeeding stage.     

Immobilization of avian excreta-derived nutrients and reduced lignin decomposition in needle and twig litter in a temperate coniferous forest

The possible effects of excreta of the Great Cormorant Phalacrocorax carbo on decomposition processes and dynamics of nutrients (N, P, Ca, K, Mg) and organic chemical components (lignin, total carbohydrates) were investigated in a temperate evergreen coniferous forest near Lake Biwa in central Japan. Two-year decomposition processes of needles and twigs of Chamaecyparis obtusa were examined at two sites, control site never colonized by the cormorants (site C) and colonizing site (site 2). Mass loss was faster in needles than in twigs. Mass loss of these litter types was faster at site C than at site 2, which was ascribed to the decreased mass loss rate of acid-insoluble ‘lignin’ at site 2. Net immobilization of N, P, and Ca occurred in needles and twigs at site 2; whereas at site C, mass of these elements decreased without immobilization during decomposition. Duration of immobilization phase of these nutrients at site 2 was estimated to be 1.6 to 2.5 years in needles and 19.6 to 23.5 years in twigs. Immobilization potential (maximum amount of exogenous nutrient immobilized per gram initial material) was similar between needles and twigs for N and Ca but was about 10 times higher in twigs than in needles for P. δ¹³C in needles was relatively constant during the first year and then increased during the second year, whereas δ¹³C in twigs was variable during decomposition. Acid-insoluble fraction was depleted in ¹³C compared to whole needles (1.6-2.1‰) and twigs (2.0-2.5‰). δ¹⁵N of needles and twigs and their acid-insoluble fractions approached to δ¹⁵N of excreta during decomposition at site 2. This result demonstrated the immobilization of excreta-derived N into litter due to the formation of acid-insoluble lignin-like substances complexed with excreta-derived N. No immobilization occurred in K and Mg and their mass decreased during decomposition at both sites. Based on these results of nutrient immobilization during decomposition and on the data of litter fall and excreta amount at site 2, we tentatively calculated stand-level immobilization potential of litter fall and its contribution to total amount of N and P deposited as excreta. Thus, the potential maximum amount immobilized into litter fall (needles and twigs) was estimated to account for 5-7% of total excreta-derived N and P.     

Soil CO2 efflux vs. soil respiration: Implications for flux models

In the long term, all CO₂ produced in the soil must be emitted by the surface and soil CO₂ efflux (FCO₂) must correspond to soil respiration (R_soil). In the short term, however, the efflux can deviate from the instantaneous soil respiration, if the amount of CO₂ stored in the soil pore-space (SCO₂) is changing. We measured FCO₂ continuously for one year using an automated chamber system. Simultaneously, vertical soil profiles of CO₂ concentration, moisture, and temperature were measured in order to assess the changes in the amount of CO₂ stored in the soil. R_soil was calculated as the sum of the rate of change of the CO₂ storage over time and FCO₂. The experiment was split into a warm and a cold season. The dependency of soil respiration and soil efflux on soil temperature and on soil moisture was analyzed separately. Only the moisture-driven model of the warm season was significantly different for FCO₂ and R_soil. At our site, a moisture-driven soil-respiration model derived from CO₂ efflux data would underestimate the importance of soil moisture. This effect can be attributed to a temporary storage of CO₂ in the soil pore-space after rainfalls where up to 40% of the respired CO₂ were stored.